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. 2017 May 27;79(7):1182–1190. doi: 10.1292/jvms.17-0084

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©2017 The Japanese Society of Veterinary Science

This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/)

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Fig. 2. — Alginate beads culture suppresses Sox-9 mRNA expression in canine bone marrow-derived mesenchymal stem cells (cBMSC). (A) Alginate encapsulated cBMSC presented in Fig. 1 were further evaluated for Sox-9 mRNA expression reverse transcriptase-PCR (RT-PCR) ethidium bromide gel electrophoresis since initial assaying by qPCR showed no detectable Sox-9 mRNA signal albeit cells expressing type II collagen and aggrecan. RT-PCR verified that Sox-9 mRNA was suppressed in cBMSC cultured in alginate beads whereas type I, II and X collagen, aggrecan and Hypoxia Inducible Factor-2α subunit (HIF-2α) mRNA bands were all detectable. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH was used as reference gene. L; 100 bp DNA ladder. Gels shown are from one representative experiment of six independent experiments (n=6), (B) shows that contrary to alginate beads culture, monolayer cBMSC cultured in CDM express Sox-9 mRNA and exhibit a chondrogenic phenotype.